Αβιοτικές καταπονήσεις στο Arabidopsis thaliana: Διερεύνηση του ρόλου των γονιδίων sbp (selenium binding protein) και ανάπτυξη ενός μοριακού εργαλίου για τον in planta ανοσοεντοπισμό τους.)

Postgraduate Thesis uoadl:1315603 550 Read counter

Unit:
ΠΜΣ Μικροβιακή Βιοτεχνολογία
Library of the School of Science
Deposit date:
2012-04-11
Year:
2012
Author:
Σκαμάκη Καλλιόπη
Supervisors info:
Επικ. Καθηγητής Ανδρέας Ρούσσης (επιβλέπων), Επικ. Καθηγητής Κοσμάς Χαραλαμπίδης, Καθηγήτρια Αμαλία Καραγκούνη-Κύρτσου
Original Title:
Αβιοτικές καταπονήσεις στο Arabidopsis thaliana: Διερεύνηση του ρόλου των γονιδίων sbp (selenium binding protein) και ανάπτυξη ενός μοριακού εργαλίου για τον in planta ανοσοεντοπισμό τους.)
Languages:
Greek
Summary:
This master thesis aimed at studying the expression of Arabidopsis thaliana sbp
genes (sbp 1/2/3) under stress conditions induced by exposure to heavy metals.
Additionally, SBP protein interactions and subcellular localization was
studied. Previous DNA microarray studies indicated that the expression levels
of sbp genes change in response to plant’s exposure to biotic and abiotic
stress. Also, plants overexpressing sbp1 gene have shown increased tolerance in
elevated selenium concentrations. During this study, A.thaliana seedlings were
exposed to different heavy metal concentrations (chromium and copper) and the
expression levels of sbp genes were evaluated using a semi-quantitative reverse
transcription-polymerase chain reaction technique. In the same conditions the
expression levels of two genes (gapdh and fba) coding for two glycolytic
enzymes, one GAPDH and one FBA, which in a previous study has been shown to
interact with AtSBP1 in vivo and in vitro, were analyzed. Additionally, AtSBP1
interaction with AtGAPDH and AtFBA and the localization of the complexes was
studied using the Bimolecular fluorescence complementation (BiFC) method.
Furthermore, to study the expression and function of AtSBP1 protein in planta
in real time, a molecular tool based on antibody fragments VHH was developed.
Using this tool the subcellular localization of AtSBP1 was determined. The
VHHs that have been used in this study derived from an antibody fragment (VHH)
library constructed after Lama glama immunization with recombinant AtSBP1
protein. AtSBP1 immunodetection in planta in real time was realized by using
selected VHHs as intrabodies. VHH-AtSBP1 complexes were traced using the
Bimolecular fluorescence complementation (BiFC) method.
Keywords:
Selenium binding protein, Arabidopsis thaliana , Abiotic stress, In planta immunolocalization, Antibody fragments VHH
Index:
Yes
Number of index pages:
4-5
Contains images:
Yes
Number of references:
167
Number of pages:
105
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